Anti-cancer compounds

ABSTRACT

The present invention provides compounds useful to inhibit tumor growth and to induce apoptosis. In general, the anti-cancer agents (ACA) are described by the formula: 
     
       
         [ACA] n -X  [Formula I] 
       
     
     wherein X is a linker group having 2-5 functional groups or is absent, n=1, and ACA is selected from the group consisting of Formula II, Formula III, Formula IV, Formula V, and Formula VI, as described herein. Other compounds described herein are defined by the Formula VII, as described herein.

The present application claims priority to U.S. Provisional Patentapplication Ser. No. 60/097,210, filed Aug. 20, 1998 and U.S.Provisional Patent application Ser. No. 60/141,169, filed Jun. 25, 1999.

GOVERNMENT RIGHTS

This invention was made in part with government support under grantnumbers NIH HL-26284, awarded by the National Institutes of Health. Thegovernment has certain rights to this invention.

FIELD OF THE INVENTION

The present invention relates generally to the field of cancertreatments, as well as to the field of peptide and non-peptidepharmaceutical compounds.

BACKGROUND OF THE INVENTION

Many lung and prostate cancers, of which small cell lung cancer (SCLC)is a prime example, have a neuroendocrine phenotype, and their growth isstimulated by neuropeptides. Antagonists of several peptides (e.g.bradykinin, substance P, bombesin) have been used in experimentaltreatment of models of SCLC in animals. Among the most potent of thepeptides examined thus far, crosslinked dimers of certain bradykininantagonist peptides have been efficacious both in vitro and in vivoagainst strains of SCLC and other tumors Chan et al., Immunopharmacology33: 201-204, 1996; Stewart et al., Can. J Physiol. Pharmacol 75:719-724, 1997; Stewart et al., U.S. Pat. No. 5,849,863, issued Dec. 15,1998). Prostate cancers show a similar neuroendocrine phenotype and aresusceptible to neuropeptide antagonists.

SUMMARY OF THE INVENTION

The present invention provides anti-cancer agents (ACA) comprised of arange of novel amino acid derivatives and small peptides having theability to inhibit growth of SCLC and certain other tumor cell lines(such as non-small cell lung cancer (NSCLC) and prostate cancer) instandard in vitro tests, as well as certain monomeric peptides that showinhibition of tumor growth in vivo. Certain of the peptides have ageneral structural relationship to carboxy-terminal fragments ofbradykinin antagonists, but the non-peptides show no such generalrelationship. Monomers, dimers, trimers, tetramers, pentamers andcyclized analogs of the novel molecules are described. The new compoundsare tested for bradykinin antagonist activity in standard assays, butthere is no apparent relationship between bradykinin antagonist activityand cytolytic potency. All of the molecules described possess bothhydrophobic (usually aromatic) and basic groups in their structures.Without being held to one particular theory, it appears that thecompounds function by stimulation of cell death (apoptosis) in the tumorcells.

The present invention also provides compounds and methods for inhibitingcancer by administering to a subject afflicted with cancer (ie. of thelung or prostate) a therapeutically effective amount of one or more ofthe compounds herein described.

In general, the anti-cancer compounds are described by the formula:

[ACA]_(n)-X  [Formula I]

wherein X is a linker having 2-5 functional groups or is absent, n=1-5,and ACA is selected from the group consisting of Formula II, FormulaIII, Formula IV, Formula V, and Formula VI. Other compounds describedherein are defined by the Formula VII. The specifics regarding structureare enumerated in the Detailed Description, Examples and Claims. Certainphysical charateristics are enumerated in the Examples as well as theDetailed Description, Examples and Claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows inhibition of growth in vivo of SCLC strain SHP-77 byB10054.

FIG. 2 shows inhibition of growth in vivo of NSCLC strain A-549 by M620.

FIG. 3 shows inhibition of growth in vivo of SCLC strain SHP-77 byB9430.

FIG. 4 shows inhibition of growth in vivo of SCLC strain SHP-77 byB10238

FIG. 5 shows inhibition of growth in vivo of SCLC strain SHP-77 by M570,both as the trifluoroacetate salt and as the hydrochloride salt.

FIG. 6 shows inhibition of growth in vivo of SCLC strain SHP-77 by M822.

FIG. 7 shows inhibition of growth in vivo of SCLC strain SHP-77 by M638.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a range of monomeric, dimeric, trimeric,tetrameric, pentameric and cyclic small peptides and peptide mimics thatare effective as anti-cancer agents.

In general, the anti-cancer agents (ACA) are described by the formula:

[ACA]_(n)-X  [Formula I]

wherein X is a linker group having 2-5 functional groups or is absent,n=1, and ACA is selected from the group consisting of Formula II,Formula III, Formula IV, Formula V, and Formula VI, as described herein.Other compounds described herein are defined by the Formula VII, asdescribed herein.

X can be any linking group which does not interfere with the inhibitoryactivity of the monomer-linker or oligomerized product usingbis-imido-esters, bis-maleimidoalkanes such as bis-maleimidohexane,dicarboxylic acids, tricarboxylic acids, tetracarboxylic acids and multicarboxylic acids. Alkane groups may be substituted with alkyl, amino,carboxyl, halogen, hydroxy, mercapto or methoxy groups. Aminoalkyl,aromatic or cycloalkyl polycarboxylic acids, heterocyclic polycarboxylicacids, carboxylic anhydrides and polyoxyethylene linkers may also beused. For C-terminal crosslinking, X may be any diamino or polyaminoalkane, cycloalkane, aromatic, heterocyclic diamine, polyamine or othersubstituted chelating agent (for example: diethylenetriaminepentaaceticdianhydride, ethylenediaminetetraacetic dianhydride, etc.).Polyamino-polycarboxylic acids may also be used to make heteromers (suchas ethylenediamine-N,N′-diacetic acid, etc.).

The linkage may be at the N-terminal or the C-terminal or at anyposition of the ACA sequence through side-chain functional groups. Thelinker may have any chain length.

For dimers, there is a correspondence between linker length andcytotoxicity. Alkyl chains of 8 carbons or more are preferred, withthose of 8 to 18 carbons being most preferred. Examples of preferreddimer linkers for the α-amino at the N-terminal or for a basicside-chain group at any position of ACA include ADA, BTAC, DDD, DDS,DTP, EGS, EOPC, HDD, HFG, PFS, SBEC, SUB, SUIM and TDIM. Fordimerization through the C-terminal carboxyl or any side-chain carboxylin ACA, the preferred linkers include DDA, DEA, EDA, EDP and HAD. Anydi-functional molecule can be used.

For trimers, linkers for basic groups include BTAC, BTC, CHTC, CTAC andTREN-(SuC)₃; for carboxyl groups, TREN. Any tri-functional molecule canbe used.

For tetramers, linkers can be BAPTA, CPTA, EDTA, EGTA, ETTA, or anytetra-functional molecule.

For pentamers, the linker can be DTPA or any pentameric functionalmolecule. Compounds formed by ACA and a linker X may be homo or heteromultimers.

[Formula II] comprises:

R-A⁻¹-B⁰-C¹-D²-E³-F⁴-G⁵-H⁶-I⁷-J⁸-K⁹

wherein R, A, B, C, D, E, F, G, H, I, J, and K are selected from thefollowing or may be absent, and wherein K is Arg or an Arg derivative:

R A⁻¹ B⁰ C¹ D² E³ F⁴ G⁵ H⁶ I⁷ J⁸ K⁹ Absent Absent Absent Absent AbsentAbsent Absent Absent Absent Absent Absent Absent or or or or Or or or oror or or or 3,3DP DmK Apc ApC MeP Hyp Ava Add Arg 2Nal Arg Aaa Lys ArgArg Nig Pro BAla Aud Gly DCpG 2Nal Arg(H) Ac Lys(εFlu) DArg DmK NMF DprCpG Pac DF5F 2Nal-NH₂ Arg-CH₂OH Aca NiK DLys NiK Pro Eac DDMF Pac DIgl3,4F2F Arg-NH₂ BApg PzO DmK NiO Gly DMF Ser DPFF 3Pal Arg(NO₂) Cca DniKPaF Eac Thr DPHe Ac6c Arg-OMe Cin DpaF PzxO Igl DTic Aic DArg Dca DPzKLys Gly Ana DArg-NH₂ Dcg DPzO Pac mABz Apb DArg(NO₂) Dhq Lys Phe pABzApb Dmac NiK Thi Pac Atpc Dpa PaF PaF(Dcg) Bip F5bz PzO pAmb Cmp F5cDArg-(NO₂) Arg-(NO₂) CpG F5pa DhPhe Gun Dpr(Fbz) Hxa Dpr(Paa) Mca F5FMcg F5F-NH₂ Moti Hphe Pcc Ica Ppa Igl Pya Igl-NH₂ Saa Ileu Ste Lys(CH₃)₃Tfmc Lys(F5bz) Mapa MBC MFF Nc6G Nc7G NMF OBS OBT OBY OC2Y Oic Oic-NH₂PABz Pac PaF(F5c) PaF(Fbz) PaF(Mcg) PaF(Ppa) PaF(Sin) pAmb pAPa PCF PdFPFF PFF-NH₂ Phe PNF Thi Tic Trp Trx Tyr

[Formula III] comprises:

R-A¹-B²-C³-D⁴-E⁵-F⁶

wherein R, A, B, C, D, E, and F are selected from the following or maybe absent, and wherein F is not Arg or an Arg derivative:

R A¹ B² C³ D⁴ E⁵ F⁶ Absent or Absent or Absent or Absent or Absent orAbsent or Absent or 2,2Dp DArg Arg Add 2Nal 1Nal 2Nal 3,3Dp DArg(NO₂)Aud 3Pal 2Nal 3Pal Aaa Ava Arg 2Nap ABza Ac Eac Arg(Tos) 3Pal ABza AcaLys Atcp Apa Ama Boc Pac D2Nal Arg Ampy Chc Darg Arg-NH₂ Ampz CinDArg(Tos) Asp Apa Ctim DF5F Atc Api Dca DIgl Atcp Aptp Dcg DPFF Bip AqdDhq Eac BtA Aqu Dmac F5F Cys(Meb) Arg(H) Dns Gly Cys(SO3H) Arg-CH₂OH DpaHis D2Nal Arg-NH₂ F5c Igl DArg Arg-OMe F5pa mABz DArg-NH₂ Asp F5po OC2YF5F Asp(Aqu) Gbc Pac Glu Atcp Gun PFF Gly Atmp Hxa Igl AtmpO Mcg InpAtpm Mse Iqa Cyh Pya mABz Dmab Seb MC2Y Dmm Sin N-Dmb-Tyr(Bz)-OMe DmpSul OC2Y Dpea Tfmc OCIY Dpma Tha Oic Dpr(Dcg-2-Nap) pABz Ecap PaF(Mes)F5F-NH₂ PFF GaP Tic mA₂Bz tLeu mA₂Bz(Dcg) Trp mA₂Bz(Gun) Try mABzTry(Bzl) Mapp Tyr Matp Arg(NO₂) MatpO pABz PaF PaF(Dcg) PaF(Mcg) PaF-NH₂PFF-NH₂ PgF PzO Sud Thm Thm Tpac Tpac Tyr(Bz)O Me

[Formula IV] comprises:

A⁰-B¹-C²-D³-E⁴-F⁵-G⁶-H⁷-I⁸-J⁹-K¹⁰-L¹¹

wherein A, B, C, D, E, F, G, H, I, J, K and L are selected from thefollowing or may be absent:

A⁰ B¹ C² D³ E⁴ F⁵ G⁶ H⁷ I⁸ J⁹ K¹⁰ L¹¹ Absent Absent Absent Absent AbsentAbsent Absent Absent Absent Absent Absent Absent or or or or or or or oror or or or DArg Arg Pro Lys Pro DTrp Gln DTrp Phe DTrp Leu(r) Leu-NH₂DArg DNMF Leu

[Formula V] comprises:

X-c[A^(−A)-B⁰-C¹-D²-E³-F⁴-G⁵-H⁶-I⁷-J⁸-K⁹]

wherein X, A, B, C, D, E, F, G, H, I, J, and K are selected from thefollowing or may be absent:

X A B C D E F G H I J K −1 0 1 2 3 4 5 6 7 8 9 Absent Absent AbsentAbsent Absent Absent Absent Absent Absent Absent Absent Absent or or oror or or or or or or or or α-Aca Ava DArg Arg Pro Hyp Gly Add DArg DDabDTrp Arg 3,3Dp BAla DNik NiK Aud Ser DDpr F5F Leu DmK DPaF PzO Ava ThrDF5F Lys NiK Glt DPzK BAla DIgl Nc7G PaF Lys DPzO DNMF DLys Oic 3Pal SucEac DOm PaF Igl DPaF PFF Thi Nig Phe Pac Phe

[Formula V] also comprises:

X-c[A⁻¹-B⁰-C¹-D²-E³-F⁴-G⁵-H⁶-I⁷-J⁸]-K⁹

[Formula V] also comprises:

X-c[A⁻¹-B⁰-C¹-D²-E³-F⁴-G⁵-H⁶-I⁷]-J⁸-K⁹

wherein the cyclization is via a side chain functional group other thanthe C-terminal residue and the residues are as described in theimmediately preceding table.

[Formula VI] comprises the following cyclic peptides:

ACA can also be those compounds in Table 4.

[Formula VII] comprises:

[ACA]₁-Eac-Eac-[ACA]₂

wherein [ACA] is defined by Formula I or the compounds in Table 4.

The in vivo inhibitory effects of antagonists may be studied usingtumor-bearing nude mice. A tumor model employing nude miceorthotopically implanted with human lung cancer cells wherein the ACA isdelivered by intratracheal instillation and aerosol inhalation may beused to evaluate the efficacy and feasibility of these antagonists as ameans of treating human lung cancers. Control animals without tumorimplantation may also be used to study the general side effects orcytotoxicity of the compounds. It is believed that aerosolized deliveryor intratracheal instillation of the agents can produce effective doseaccumulation in the area of lesion and reduce the overall systemictoxicity of the compounds in the animals more than when the compound isdelivered by systemic administration.

The compounds may be administered topically, or by injection or infusionor as an oral suspension in an appropriate vehicle or as tablets, pills,capsules, caplets or the like, or preferably via intratrachealinstillation or aerosol inhalation. The dosage and manner ofadministration will be defined by the application of the ACA and can bedetermined by routine methods of clinical testing to find the optimumdose. These doses are expected to be in the range of 0.001 mg/Kg to 100mg/Kg of active compound.

The compounds are composed of amino acids which may form salts due totheir acidic or basic nature, and any pharmacologically acceptable saltderived from the compounds described in this invention such ashydrochlorides, acetates, phosphates, maleates, citrates, benzoates,salicylates, succinates, ascorbates and the like, including HCl,trifluoroacetic acid (TFA), and HOAc, are considered an extension ofthis invention. A common tactic in medicinal chemistry is to modifyknown drug substances which are peptide based to form esters or amideswhich exhibit greater bioavailability. Prodrugs derived from thecompounds disclosed here are therefore considered an obvious extensionof this invention. Methods for designing and preparing prodrugs aredescribed in detail in the medicinal chemical literature.

Structures and biological activities of peptides and peptide mimicsrelated to bradykinin (BKR) are given in Table 1. Structures andbiological activities of compounds not related to bradykinin (BKU) aregiven in Table 2. Structures and biological activities of cyclicpeptides are given in Table 3. Structures of previously described knownpeptides which we have found to be active against cancers in vivo areincluded in Table 4. Actions of selected compounds on prostate cancercell lines are given in Table 5. Abbreviations used are as defined inTable 6.

EXAMPLES

In general, Anti-bradykinin activity was determined by the classicalguinea pig ileum assay and on Chinese hamster ovary (CHO) cellsexpressing cloned human bradykinin B2 receptors. Anti-tumor activity wasdetermined on cultured human cancer cell lines using the standardtetrazolium (MTT) assay. No correlation between anti-bradykinin andcytolytic activity was found among the compounds, indicating that cellsare not killed due to inhibition of an essential bradykinin function.Potent compounds were found to stimulate apoptosis in SCLC cells,probably by abnormal activation of the intracellular MEKK pathway.

EXAMPLE I

Synthesis of Peptides

Peptides were synthesized using standard solid phase synthesis methods,well known in the art (Stewart and Young, Solid Phase Peptide Synthesis,Pierce Chemical Co., Rockford, Ill., 1984) and were purified by HPLC andwere characterized by amino acid analysis (AAA), thin layerchromatography (TLC) and laser desorption mass spectrometry (LDMS).Peptide amides were synthesized on methylbenzhydrylamine (MBHA) resin,which yields amides directly. Peptide methyl esters (OMe) weresynthesized by reaction of peptides with 2,2-dimethoxypropane (Rachele,J. Org. Chem. 28: 2898, 1963). Cyclic peptides were prepared on resin orin solution with PyAOP and HOAt.

EXAMPLE II

Synthesis of Non-peptides

Non-peptides were synthesized by standard organic chemistry procedureswell known in the art. Compounds were purified by HPLC and werecharacterized by analytical HPLC, TLC, and LDMS.

EXAMPLE III

Synthesis of DDD and SUB Dimers

Synthesis on Resin

Neutralized peptide-resin (0.05 mmole) was treated with 0.15 mmolediisopropylethyl amine (DIEA) and 0.026 mmole dodecanedioyl dichlorideor suberoyl dichloride in 2.5 mL dichloromethane (DCM). The suspensionwas mixed for 5 h, washed with DCM and ethanol and dried. The peptidedimer was cleaved from the resin with HF, and the peptide was extractedand purified

Synthesis in Solution

Carboxyl-derivatized amino acids or dipeptides were dissolved indimethyl formamide (DMF) and treated with 10 equivalents of DIEA and0.55 equivalent of dodecanedioyl dichloride or suberoyl dichlorideovernight. The DMF was evaporated in vacuo and the resulting dimer waspurified by HPLC.

EXAMPLE IV

Synthesis of EGS, DTP, SBEC and SUB Dimers in Solution

Dimerization in solution proceeded by reacting 1 equivalent of peptidemonomer trifluoroacetate, an excess of DIEA and 0.55 equivalent ofcross-linking reagent overnight in DMF. The cross-linking agents werepurchased from Pierce (EGS dimer, ethylene glycolbis-(succinimidylsuccinate); DTP dimer, dithiobis (succinimidylpropionate); SBEC dimer, bis[(2(succinimidooxycarbonyloxy)ethyl]sulfone;SUB dimer, disuccinimidyl suberate).

EXAMPLE V

Synthesis of Boc-N-cycloheptylglycine (Nc7G)

N-Cycloheptylglycine was synthesized by reductive amination ofcycloheptanone with glycine methyl ester following the proceduredescribed in Gera et al., Immunopharmacology. 33:174-177 (1996). Thecrude product was converted to the N-Boc derivative (mp, 89-90° C.).

EXAMPLE VI

Synthesis of TDIM Dimers

Dimethyl tetradecyldiimidate was synthesized from tetradecanedinitrileby the method of De Abreu et al. (Eur. J Biochem. 97: 379-387, 1979. Oneequivalent of peptide TFA salt or other molecule having a free aminogroup was dissolved in DMF and stirred with 10 equivalents of DIEA and0.7 equivalent of dimethyl tetradecyldiimidate dihydrochloride overnightat room temperature. DMF was evaporated in vacuo and the dimer waspurified. SUIM dimers were prepared similarly, using dimethylsuberimidate.

EXAMPLE VII

Synthesis of B10238: F5C-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg(F5c-B9430)

B10238 was made by standard solid phase synthesis procedures, or by theacylation of B9430 with 2,3,4,5,6-pentafluorocinnamic acid in DMF, usingBOP coupling in presence of excess DIEA. The product was purified byHPLC.

EXAMPLE VIII

Synthesis of M822: DDD-(DArg-F5F-Arg)₂

Following standard solid phase synthesis procedures, Boc-Arg(Tos)Merrifield synthesis resin was coupled with Boc-F5F, followed bycoupling with Boc-DArg(Tos), using HATU as coupling agent. Thepeptide-resin was deprotected with TFA-DCM and neutralized with TEA. Thepeptide-resin was then treated with 0.55 equivalent of dodecanedioyldichloride and 5 equivalents of DIEA in DCM overnight at roomtemperature. After washing and drying, the resin was cleaved withanhydrous HF, using standard conditions. The peptide was extracted fromthe resin with 90% acetic acid and purified by preparative HPLC.

EXAMPLE IX

Synthesis of M570 Hydrochloride: F5c-OC2Y-Atmp.HCl

4-Amino-2,2,6,6-tetramethylpiperidine (Aldrich) was coupled withBoc-(O-2,6-dichlorobenzyl)-tyrosine, using BOP in DMF solution. The Bocprotecting group was removed by TFA and the product coupled with2,3,4,5,6-pentafluorocinnamic acid in DMF, using BOP in the presence ofexcess DIEA at room temperature for 3 h. The DMF was removed in vacuo,the product was extracted into ethyl acetate and the solvent wasevaporated. The residue was treated with 0.1-1.0 N HCl or 20% ethanolicHCl. The solvent was removed by evaporation in vacuo at roomtemperature. The residue was lyophilized from water-dioxane orcrystallized from ethanol-ether.

EXAMPLE X

Synthesis of M630: Dmac-OC2Y-Matp.TFA

4-Methylamino-2,2,6,6-tetramethylpiperidine (Matp) was synthesized from2,2,6,6-tetramethyl-4-piperidone (Aldrich) and methylamine by reductiveamination with NaCNBH₃. The Matp was coupled withBoc-(O-2,6-dichlorobenzyl)-tyrosine, using BOP in DMF solution. The Bocprotecting group was removed by TFA and the product was coupled with4-(dimethylamino)cinnamic acid in DMF, using BOP in the presence ofexcess DIEA at room temperature for 3 h. The DMF was removed in vacuo.The product was extracted into ethyl acetate and the solvent wasevaporated in vacuo. The crude product was purified by HPLC, giving theTFA salt. The Dmac-OCTY-Matp.TFA salt can be converted to its HCL saltas in Example IX above.

EXAMPLE XI

Synthesis of M638: DDD-(DArg-Igl-Arg-Matp)₂

In sequence, Boc-Arg(Tos), Boc-Igl and Boc-DArg(Tos) were coupled to4-methylamino-2,2,6,6-tetramethylpiperidine (Matp), using BOP ascoupling agent in DMF in the presence of excess DIEA at room temperaturefor 3-5 h. After removal of DMF in vacuo, the product was extracted intoethyl acetate. After evaporation of the solvent, the residue was treatedwith TFA-DCM to remove the Boc group. TFA was removed in vacuo. TheDArg(Tos)-Igl-Arg(Tos)-Matp.TFA was treated with dodecanedioyldichloride (0.55 equiv) and DIEA (5 equiv) in DCM for 5 h. Theprotecting groups were cleaved by HF and the lyophilized product waspurified by HPLC. The M638.TFA salt was converted to its HCl salt, using0.1-1.0 N HCl or 20% ethanolic HCl as in Example IX above.

EXAMPLE XII

Synthesis of M590: Atmp-Igl-Pac-α-Sbl-Lys-B9430

In sequence, Boc-Igl, Boc-Pac and mono-methyl sebacate were coupled to4-amino-2,2,6,6-tetramethylpiperidine (Atmp), using BOP coupling agentin DMF in presence of excess DIEA at room temperature for 3-5 h. DMF wasremoved in vacuo and the product was extracted into ethyl acetate. Afterevaporation of the solvent, the methyl ester was hydrolyzed in methanolby 1N NaOH. The crude product (0.025 mmol Atmp-Igl-Pac-Sbl) was coupledto the peptide resin (0.02 mmolLys(2-CIZ)-DArg(Tos)-Arg(Tos)-Pro-Hyp-Gly-Igl-Ser(Bzl)-DIgl-Oic-Arg(Tos)-Merrifieldresin) using BOP/DIEA activation in DMF. The heterodimer peptide wascleaved from the resin with HF, using standard conditions. The peptidewas extracted from the resin with acetic acid and purified bypreparative HPLC.

EXAMPLE XIII

Synthesis of M872: c[DArg-Arg-Eac-Ser-DF5F-Oic-Arg]

Following standard solid phase synthesis procedures, Boc-DArg(Tos) wascoupled to Boc-Arg(Tos) Merrifield synthesis resin, followed in sequenceby Boc-Arg(Tos), Boc-Oic, Boc-DF5F, Boc-Ser(Bzl), and Boc-Eac, usingHATU as coupling agent. After deprotection with TFA-DCM, the resin wascleaved with anhydrous HF using standard conditions. The peptide wasextracted from the resin with 0.1% TFA-H₂O/dioxane and lyophilized. Thepeptide trifluoroacetate was cyclized with three equivalents of PyAOPand HOAt and 20 equivalents of DIEA in DMF at a concentration of 10⁻³ M.After removal of the solvent under reduced pressure, the product waslyophilized from dioxane-H₂O and purified by HPLC.

EXAMPLE XIV

Synthesis of M678: (Dns-DArg-Igl-Arg)₂-DDA

In sequence, Boc-Arg(Tos), Boc-Igl and Boc-DArg(Tos) (2 equivalents)were coupled to 1,10-decanediamine using BOP as a coupling agent in DMFin presence of excess DIEA at room temperature for 3-5 h. DMF wasremoved in vacuo and the product was extracted into ethyl acetate. Thesolvent was evaporated in vacuo and the residue was treated with TFA/DCMto remove the Boc group. TFA was removed in vacuo, and the product wastreated with dansyl chloride (2 equivalents) and an excess of DIEA inDCM for 5 h. The Tos groups were cleaved by HF and the crude product waspurified by HPLC.

EXAMPLE XV

Synthesis of M290: BTAC-(2-Nal-Atmp)₃

The benzene-1,3,5-tris-carbamido-ε-caproic acid linker was made from1,3,5-benzenetricarboxylic acid and N-Boc-ε-caproic acid methyl ester,using the BOP coupling method. The methyl ester was hydrolyzed inmethanol by 1N NaOH. The product (1 equivalent BTAC) was coupled to2-Nal-Atmp (3 equivalents) in DMF, using HATU as coupling agent. Thesolvent was removed in vacuo, and the residue was purified by HPLC. TheBTAC-(2-Nal-Atmp)₂-OH was also isolated as a by-product.

EXAMPLE XVI

Synthesis of M1040: EDTA-(OC2Y-ATMP)₄

Boc-(O-2,6-dichlorobenzyl)-tyrosine was coupled with4-amino-2,2,6,6-tetramethylpiperidine overnight in DMF, using BOP ascoupling agent in the presence of DIEA. After removal of DMF in vacuo,the residue was extracted into ethyl acetate and treated with TFA/DCM tocleave the Boc group. The TFA/DCM was evaporated in vacuo and theproduct (OCTY-ATMP) was lyophilized from dioxane/water.Ethylenediaminetetraacetic acid (0.25 equivalent EDTA) was coupled withOC2Y-ATMP trifluoroacetate (1 equivalent) in DMF, using BOP as couplingagent in the presence of DIEA. The solvent was removed in vacuo and theresidue was purified by HPLC.

EXAMPLE XVII

Assay of Anti-bradykinin Activity on Guinea Pig Ileum

Male Hartley guinea pigs that had been deprived of food overnight weresacrificed, and sections of terminal ileum, 25 mm in length, weredissected, attached to tissue holders and immersed in 10 ml tissue bathscontaining Krebs' solution bubbled with 95%O₂/5%CO₂. Tissues were placedunder 1 g tension and incubated for 1 h equilibration.Concentration-effect curves were constructed to bradykinin in theabsence and presence of new compounds. Bradykinin showed pD₂=7.4, andantagonist B9430 showed pA₂=7.9.

EXAMPLE XVIII

Assay of Anti-bradykinin Activity on Cloned Human B2 Receptors

Chinese hamster ovary cells containing cloned and expressed humanbradykinin B2 receptors were grown in cell cups of the Cytosensormicrophysiometer in Ham's F-12 medium supplemented with sodium pyruvateand 10% FBS (Gibco 11765-054). For assay the cells were transferred toHam's F-12 without bicarbonate or serum (Gibco 21700-075) and placed inthe Cytosensor. Concentration-response curves were constructed tobradykinin in the presence or absence of new compounds. Bradykininshowed pD₂=11, and antagonist B9430 showed pA₂=10.5

EXAMPLE XIX

Colorimetric Tetrazolium Assay for Cell Survival

Cell growth and survival were measured by a rapid colorimetric assaybased on the tetrazolium salt MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)(Mosmann, J Immunol. Methods 65: 55-63, 1983, with minor modifications).Briefly, 1,000 normal lung fibroblasts or normal epithelial BEAS-2Bcells, 1,000 or 5,000 viable non-SCLC cells or 10,000 viable SCLC cellswere plated in 100 μL of growth medium in 96-well flat-bottomedmicrotiter plates. Cells were incubated overnight to allow recovery.Compounds to be tested were added to the cells in triplicate in a rangeof concentrations and the cells were incubated at 37∞C, 5% CO₂, with100% humidity. Control cells were treated in the same way withoutantagonists. All wells had a final volume of 200 μL. Plates wereincubated for 4 days, allowing sufficient time for cell replication andcompound-induced cell death to occur. On day 5, 25 μL of a 2 mg/mLsolution of MTT (Sigma) dissolved in RMPI-1640 was added to each well.The plate was incubated for 4 h at 37∞C. The supernate was removed andthe blue formazan complex was dissolved by adding 100 μL of 0.02 N HClin 75% isopropanol to all wells. Absorbance was immediately determinedusing a scanning multiwell plate reader. B9870 caused 50% cell death ata concentration of 0.15 μM under these conditions.

EXAMPLE XX

Measurement of Apoptosis in Cultured Cells

Apoptosis, also known as programmed cell death, is the phenomenon bywhich a cell dies following a series of gene-mediated events, inresponse to a wide range of intracellular and extracellular agents.Apoptosis, a counterpart of mitosis, plays an important role in thedevelopment and homeostasis of many organisms and tissues. It serves toregulate cell numbers, to shape developing organisms and as a defenseagainst potentially harmful agents. Apoptosis is not the only mode ofcell death. Necrosis is a type of cell death which is nonspecific andfrequently occurs when cells are exposed to high doses of toxic agents.Such exposure usually results in the loss of ionic homeostasis. Unlikeapoptosis, necrosis does not seem to be genetically influenced.

Apoptotic and necrotic cells have different appearances which allow themto be distinguished microscopically. Necrotic cells and theirmitochondria swell, the cell membrane eventually ruptures, and internalorganelles become distended. As a result of the membrane rupture,inflammation occurs in the surrounding tissue. In contrast, the nucleiof apoptotic cells become fragmented into several smaller nuclearbodies, which are quickly recognized by phagocytes and engulfed, and noinflammatory response occurs. Therefore, it is useful to developchemotherapeutics which induce apoptosis, rather than necrosis, in orderto avoid inflammation and the toxic agents which are often released fromnecrotic tumor cells.

We have used differential fluorescent dye uptake and cellular morphologyto distinguish viable and dead cells with apoptotic and/or necroticmorphologies. We have used Rhodamine 123 to stain active mitochondria inviable cells, Hoechst 33324 to stain DNA in both viable and dead cells,and Propidium Iodide to stain DNA in dead cells. These cellsubpopulations may be distinguished by the different manners in whichthey take up the fluorescent probes. The dead apoptotic and necroticsubpopulation, which has lost its membrane potential and organellefunction, takes up Propidium Iodide and Hoechst 33324. Since the cellsin this subpopulation are dead, the mitochondria are not active and thusthere is little or no uptake of Rhodamine 123. Under the fluorescencemicroscope with a DAPI filter, nuclei in these cells appear pinkish incolor due to the mixing of both Propidium Iodide and Hoechst 33324 dyes.Necrotic cells have intact nuclei while apoptotic cells have fragmentedmulti-nucleated bodies. In contrast, the viable apoptotic subpopulationhas an intact membrane but inactive mitochondria. As a result, thefragmented multi-nucleated bodies (a hallmark of apoptotic cells) inthese cells take up only Hoechst 33324, which gives them a blueappearance under the fluorescence microscope, but are unable to take upPropidium Iodide or Rhodamine 123. The subpopulation of viable cells hasboth intact cell membranes and active mitochondria. These cells take upboth Hoechst 3324 and Rhodamine 123. Microscopically these cells appearto have single normal blue nuclei when examined with a DAPI filter andbright green mitochondria when examined with a FITC filter.

EXAMPLE XXI

Inhibition of Tumor Growth In vivo in Nude Mice

Representative peptide and non-peptide compounds having high in vitrocytotoxic activity were tested against implanted tumors in vivo. Athymicnude mice were implanted subcutaneously with either single cellsuspensions (2 million SCLC cells or 1 million NSCLC cells) or withsmall fragments (3×3 mm) of tumors minced from previously grown nudemouse heterotransplants. On the seventh day after tumor implantationgroups of 5 mice bearing implants were injected intraperitoneally dailywith the compounds being tested at 1, 5, or 10 mg/kg/day; controlanimals were injected with an equal volume of isotonic saline. Tumorsize was measured with a caliper three times per week. Tumor volume wascalculated by the formula:

Volume(cc)=πx(length)x(width)²/6

Results of representative in vivo tests are given in FIGS. 1-8. Forcomparison, bradykinin antagonist peptide dimers B9870 and B10054 causedmarked inhibition of growth of the SCLC line SHP-77 at a dose of 5mg/kg/day.

EXAMPLE XXII

Data

Examples of peptides and peptide mimics related to the C-terminal partof bradykinin antagonist peptides and their biological activities oncancer cells and bradykinin responses are given in Table 1.

Many compounds not directly related to the structure of bradykinin weresynthesized and tested for anti-tumor and anti-bradykinin activity.These are listed in Table 2.

Cyclic peptides related to bradykinin and bradykinin mimics are reportedin Table 3, along with their biological activity on cancer cells andanti-bradykinin activity.

Structures of previously described known peptides which have been foundto be active against cancers in vivo are included in Table 4.

Cytotoxic activity in vitro of compounds M570 and M590 against variousstandard strains of prostate cancer is reported in Table 5.

Standard abbreviations were used for natural amino acids. Fornon-natural amino acids, derivatizing groups and other chemicals, theabbreviations listed in Table 6 are used.

TABLE 1 ACTIVITIES OF PEPTIDES RELATED TO BRADYKININ STRUCTURE NUMBERSTRUCTURE MTT^(a) GPI^(b) HUMAN^(c) BK^(d)Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg B9430^(d)DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg 120 8.2 B9870-2^(d)SUIM-(DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic 0.15 8.4 Arg) B8838DArg-Arg-Pro-Hyp-Gly-CpG-Ser-DCpG-CpG-Arg — 7.0 B8840DArg-Arg-Pro-Hyp-Gly-Phe-Ser-DCpG-CpG-Arg — 6.8 B8858DArg-Arg-Pro-Hyp-Gly-Thi-Ser-CpG-DCpG-DArg — 5.2 B8994DArg-Arg-Pro-MeP-Gly-CpG-Ser-DCpG-CpG-Arg — — B9074Dhq-DArg-Arg-Pro-Hyp-Gly-CpG-Ser-DCpG-CpG- — 6.3 Arg B9126Aaa-DArg-Arg-Pro-Hyp-Gly-(D,L)DMF-Ser-DTic- — 6.4 Oic-Arg B9126-2Aaa-DArg-Arg-Pro-Hyp-Gly-(D,L)DMF-Ser-DTic- — 7.3 Oic-Arg B9224-2Aca-DArg-Arg-Pro-Hyp-Gly-Thi-Ser-(D,L)Igl-Oic- — 8.4 Arg B9882

— — B9914 Oic-Arg — Wk B9916 DIgl-Oic-Arg — Wk B9490Dcg-Digl-Oic-Arg >60 Wk B9918 Ser-Digl-Oic-Arg — — B9920Igl-Ser-DIgl-Oic-Arg — — B9922 Gly-Igl-Ser-DIgl-Oic-Arg — Wk B9924Hyp-Gly-Igl-Ser-Digl-Oic-Arg — — B9926 Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg— — B9950

8 — B9956 α-DDD-(Lys-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl- — — Oic-Arg)₂.B9960 DArg-Arg-Nig-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg — 7.7 B9966DArg-Arg-NMF-Hyp-Gly-Thi-Ser-DIgl-Oic-Arg — 6.9 B10010DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Nc7G-Arg — 7.7 B10014DArg-Arg-Pro-Hyp-Gly-Thi-Ser-DTic-Nc6G-Arg — 7.6 B10054DDD-(Lys-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic- 0.3 7.1 Arg)₂ B10062DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg-NH₂ Inact 7.1 B10082SUIM-(DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg- 0.7 7.2 NH₂)₂ B10084BApG-DArg-Arg-Pro-Ryp-Gly-Igl-Ser-DIgl-Oic-Arg >20 8.1 B10088DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg-Eac- 4 7.1Eac-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg B10092(Gun)₂-BApG-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl- >20 8.7 Oic-Arg B10098(DArg-Arg-Pro-Hyp)₂-Dpr-Igl-Ser-DIgl-Oic-Arg 20 5.3 B10100-2TDIM-(DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic- 1 8.0 Arg)₂ B10100-1Moti-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg 4 7.8 B10104-2TDIM-(DArg-Arg-Pro-Hyp-Gly-Thi-Ser-DIgl-Oic- 4 8.0 Arg)₂ B10104-3Moti-Darg-Arg-Pro-Hyp-Gly-Thi-Ser-DIGl-Oic-Arg 20 8.1 B10160Leu-DTrp-Phe-DTrp-DNMF-Eac₂-DArg-Arg-Pro- 10 6.3Hyp-Gly-Igl-Ser-DIgl-Oic-Arg B10162Leu-Leu-DTrp-Phe-DTrp-DNMF-Eac₂-DArg-Arg- 7 6.3Pro-Ryp-Gly-Igl-Ser-Digl-Oic-Arg B10198-1 DDD-(Eac-Arg-Digl-Oic-Arg)₂ —5.7 B10198-2 DDD-(Eac-Arg-Digl-Oic-Arg)₂ 15 — B10200DDD-(Eac-Eac-Arg-DIgl-Oic-Arg)₂ 16 5.8 B10238F5c-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgI-Oic-Arg 150 8.1 B10252EGS-(DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg)₂ 25 8.3 B10282Arg-Pro-Pro-Gly-Phe-Thr-DTic-Oic-Arg — 7.3 B10284Arg-Pro-Pro-Gly-Phe-Thr-DTic-Oic-NH₂ — 7.7 B10382DArg-PzO-Pro-Hyp-Gly-Igl-Ser-DF5F-Oic-Arg — — B10384DNiK-PzO-Pro-Hyp-Gly-Igl-Ser-DF5F-Oic-Arg — — B10386DDD-(DmK-PzO-Pro-Hyp-Gly-Igl-Ser-DF5F-Oic- — — Arg)₂ B10388DNiK-PzO-Pro-Hyp-Gly-Igl-Ser-DF5F-Oic-Arg — — B10390DNiK-PzO-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg — — B10392DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-PFF-Arg — — B10394F5c-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DF5F-PFF-Arg — — B10396F5c-DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-PFF-Arg — — M2 Dcg-D-2-Nal-Arg — —M10 Dcg-2Nal-Arg — 4.8 M20 Gun-2-Nal-Arg — 4.6 M42 Gun-Eac-DIgl-Oic-Arg— 5.0 M68 Dcg-BtA-Arg — 4.9 M70 Dcg-Igl-Arg — 4.8 M78 Dcg-Apa-Arg — 5.6M84 Dcg-Ile-Arg — 5.1 M86 Dcg-Ac6c-Arg — 5.0 M88 Gun-Ica-Arg — 4.7 M94Dcg-Aic-Arg — Wk M96 Dcg-(D,L)Atc-Arg — 4.7 M118-1 Ac-PaF(Mcg)-Arg — 4.9M118-3 Ac-PdF-Arg — 5.4 M124 Dcg-Gly-Cmp-Arg — 4.7 M128 Dcg-Gly-Oic-Arg— Wk M130 Dcg-F5F-Arg 20 4.8 M132 F5bz-F5F-Arg 60 Wk M134 Dcg-Trx-Arg —4.9 M142 Ac-PaF(Sin)-Arg — Wk M146-1 Ac-PaF(Mcg)-p-ABz-Arg — InactM146-2 Ac-PaF(Dcg)-p-ABz-Arg — 4.9 M148 F5c-p-ABz-Arg — 5.1 M160Ste-2-Nal-Arg — Wk M176 F5c-pABz-2Nal-Arg Inact 5.4 — M196FSc-Gly-mABz-2Nal-Arg Inact 5.1 — M198 Ac-Pac-Gly-m-Abz-2-Nal-Arg — 5.1M200-1 Mcg-Pac-Gly-m-ABz-2-Nal-Arg — Inact M200-2Dcg-Pac-Gly-m-ABz-2-Nal-Arg — 4.9 M216 FSc-p-APa-Arg >180 M226DDD-(Arg-DIgl-Oic-Arg)₂ 35 5.7 Inact M232-1 Dcg-Atpc-Arg — 4.7 M232-3Dcg-2-Nal-Atpc-Arg — 5.1 M346 Dcg-p-Amb-Arg — 4.6 M348 F5c-p-Amb-Arg —4.7 M352 F5c-p-Amb-APa-Arg — 4.7 M370 F5c-Arg — 4.8 M372 F5c-APb-Arg —4.6 M374 Tfmc-Arg — 4.6 M380 F5c-Tyr-Arg — Inact M382 F5c-Tic-Arg — 4.7M388 F5c-Lys{(CH₃)₃}-Arg — 4.9 M392 F5c-Ana-Arg — 4.5 M394 F5c-Bip-Arg —4.7 M398 F5c-Pac-Arg — Inact M400 DDD-(pABz-2Nal-Arg)₂ 22 5.1 11.5 M406Arg-Eac-DIgl-Ana-Arg — Inact M410 F5c-Phe-Arg — 5.4 M412 F5c-m-APa-Arg —5.8 M416 F5c-3-Pal-Arg — Wk M420 F5c-hPhe-Arg 60 7.0 10.1 M424F5c-Thi-Arg — 4.6 M426 F5c-Trp-Arg — Inact M442 F5c-Oic-Arg — — M446F5c-2Nal-Arg 60 4.7 9 M450 F5c-2Nal-Arg-NH₂ 26 4.9 Inact M484DDD-(Pac-2Nal-Arg)₂ 25 Inact Inact M494 DDD-(Lys-Pac-Gly-mABz-2Nal-NH₂)₂33 5.1 Inact M498 DDD-(Pac-2Nal-Arg-NH₂)₂ 24 4.9 Inact M500DDD-(pABz-2Nal-Arg-NH₂)₂ 40 0 11.4 M504 DDD-(Pac-2Nal-DArg-NH₂)₂ 11 5.4Wk M508 DDD-(DArg-2Nal-Arg)₂ 23 Inact — M510 DDD-(DArg-2Nal-Arg-NH₂)₂ 8Inact 11 M512 F5c-OC2Y-Arg 70 5.7 11 M516DDD-(DArg-Arg-Aud-Pac-2Nal-Arg)₂ 1.4 0 Ag M518 DDD-(DArg-OC2Y-Arg)₂ 15Wk 10 M520 F5c-OBS-Arg Inact 6.1 7 M528 F5c-MBC-Arg Inact Inact M540Pya-hPhe-Arg >100 Wk M542 Dca-hPhe-Arg 80 Wk M550 F5c-OBT-Arg 80 InactAg M552 DDD-(p-ABz-hPhe-Arg)₂ Inact Inact M554 DDD-(DArg-hPhe-Arg)₂ —5.1 M558 Dcg-hPhe-Arg 100 Wk M560 DDD-(DArg-hPhe-Arg-NH₂)₂ 50 Wk M564DDD-(DArg-OBS-Arg)₂ — Wk M590 Atmp-Igl-Pac-α-Sbl-Lys-B9430 4.5 7.5 InactM598 DDD-(Arg-DIgl-Oic-Arg-OMe)₂ 13 — 10 M600 α-DDD-(Lys-B9430-OMe)₂ 1.26.4 Ag M608 DDD-(Eac-Arg-DIgl-Oic-Arg-OMe)₂ 13 — Inact M612F5c-DArg-hPhe-Arg Inact Wk M676 DDD-(DArg-Arg-Eac-Ser-DF5F-Nc7G-Arg)₂ —— M682 F5c-Lys(F5bz)-Arg — 4.9 M686 F5c-NMF-Arg 29 5.6 M688F5c-Dpr(Fbz)-Arg — Inact M690 F5c-Dpr(Paa)-Arg — 4.8 M692DDD-(DArg-Arg-Aud-Pac-hPhe-Arg)₂ — Wk M696 F5c-DArg-Eac-2Nal-Arg Inact5.1 — M698 F5c-DArg-Arg-Aud-Pac-2Nal-Arg 7.1 Wk — M706 Cin-hPhe-ArgInact Wk M708 Ppa-hPhe-Arg Inact 5.8 M710DDD-(DArg-Arg-Aud-Pac-2Nal-DArg-NH₂)₂ 1.7 Wk M714 F5c-PCF-Arg Inact WkM718 F5c-PFF-Arg 9 5.1 M720 F5c-PaF(Ppa)-Arg Inact 4.8 M726D-Arg-Arg-Aud-PaF(F5c)-Arg Inact 5.6 M728DDD-(DArg-Arg-Aud-PaF(F5c)-Arg)₂ 4 5.3 M730 F5c-DhPhe-Arg — 4.7 M732F5c-PNF-Arg — 4.9 M734 DDD-(DArg-Arg-Aud-Pac-PaF(Fbz)-Arg)₂ 1.8 5.3 M738F5c-DArg-Eac-hPhe-Arg Inact 5.3 M746 DDD-(Pac-hPhe-Arg)₂ Inact InactM752-2 Pac-hPhe-Arg Inact Wk M752-5 Aaa-Ser-Pac-hPhe-Arg Inact Wk M752-6Aaa-Pac-hPhe-Arg Inact Wk M754 Aaa-DPhe-hPhe-Arg Inact 4.6 M756DDD-(DPhe-hPhe-Arg)₂ 18 5.2 M758 Saa-hPhe-Arg — Wk M764Aaa-DTic-hPhe-Arg — 4.8 M766 F5c-DArg-Arg-Aud-DTic-hPhe-Arg — 5.1 M770DDD-(DArg-Arg-Aud-DTic-hPhe-Arg)₂ 8 Inact M772 Aaa-DIgl-hPhe-Arg — 4.9M774 F5c-DArg-Arg-Aud-DIgl-hPhe-Arg 8 Inact M776 DDD-(DIgl-hPhe-Arg)₂ 305 M778-1 Pcc-hPhe-Arg — Wk M780 Mca-hPhe-Arg — Wk M782 Cca-hPhe-Arg — WkM784 Ac-OC2Y-Arg Inact Wk M786 DDD-(DArg-Arg-Aud-DIgl-hPhe-Arg)₂ 3.2 WkM788 F5c-DArg-Arg-Aud-DTic-Oic-Arg 9 5 M790DDD-(DArg-Arg-Aud-DTic-Oic-Arg)₂ 1.7 Inact M792F5c-DArg-Arg-Eac-Ser-DTic-Oic-Arg >100 4.9 M794DDD-(DArg-Arg-Eac-Ser-DTic-Oic-Arg)₂ 21 Inact M796F5c-DArg-Arg-Eac-Ser-DF5F-Oic-Arg 31 6.5 M802 F5c-Lys-Ser-DF5F-Oic-ArgInact 6.3 M804 DDD-(DArg-Arg-Eac-Ser-DF5F-Oic-Arg)₂ 7.3 7.7 M806Ava-Igl-Ser-DF5F-Oic-Arg Inact 5.6 M808 DDD-(Lys-Ser-DF5F-Oic-Arg)₂ 306.9 M810 F5c-F5F-Arg 40 4.6 M812 F5c-PFF-Arg-NH₂ 15 Wk M814 Ppa-PFF-ArgInact Wk M816 Dpa-PFF-Arg 52 4.6 M818 DDD-(DArg-PFF-Arg-NH₂)₂ 60 Wk M820DDD-(DArg-PFF-Arg)₂ 43 Inact M822 DDD-(DArg-F5F-Arg)₂ 25 Mixed M826F5c-MFF-Arg 76 Inact M828 F5c-3,4F2F-Arg — Wk M838F5c-DArg-Arg-Aud-DIgl-PFF-Arg 7.4 5.1 M842DDD-(DArg-Arg-Aud-DIgl-PFF-Arg)₂ 1.4 Inact M844DArg-Arg-Aud-DIgl-PFF-Arg 12 Wk M846 DDD-(DArg-Arg-Aud-DF5F-Oic-Arg)₂ 27.1 M852 F5c-DArg-Arg-Eac-Ser-DIgl-Oic-Arg Inact 5.9 M854DDD-(DArg-Arg-Eac-Ser-DIgl-Oic-Arg)₂ 7.3 5.9 M856F5c-DArg-Arg-Aud-Ser-DIgl-Oic-Arg 21 5.4 M858DDD-(DArg-Arg-Aud-Ser-DIgl-Oic-Arg)₂ 4 6.3 M860F5c-DArg-Arg-Add-Ser-DIgl-Oic-Arg 6 5.4 M862DDD-(DArg-Arg-Add-Ser-DIgl-Oic-Arg)₂ 1.3 5.6 M864DDD-(DArg-Arg-Add-Ser-DIgl-PFF-Arg)₂ 1.8 Inact M868Ac-Darg-Arg-Aud-DF5F-Oic-Arg 55 6.5 M888F5c-DArg-Arg-Aud-Ser-DF5F-Oic-Arg 12.5 6.6 M890DDD-(DArg-Arg-Aud-Ser-DF5F-Oic-Arg)₂ 1.7 5.5 M922DDD-(DNiK-Arg-Eac-Ser-DF5F-Oic-Arg)₂ — — M926 ζ-SUB-(ApC-F5F-Arg)₂ Inact— M930 α-DDD-(ApC-F5F-Arg)₂ Inact — M932DDD-(DArg-Arg-Eac-Ser-DIgl-PFF-Arg)₂ 6.0 — M936DDD-(DNiK-PzO-Eac-Ser-DF5F-Oic-Arg)₂ — — M944DDD-(DArg-Arg-Eac-Ser-DF5F-PFF-Arg)₂ 6.7 — M946F5c-DArg-Arg-Eac-Ser-DF5F-PFF-Arg — — M950α-DDD-(K-DArg-Arg-Eac-Ser-DF5F-Oic-Arg)₂ 6.7 — M952DDD-(DmK-DArg-Arg-Eac-Ser-DF5F-Oic-Arg)₂ — — M954Aaa-DArg-Arg-Eac-Ser-DF5F-Oic-Arg 10 — M956Aaa-DArg-Arg-Aud-Ser-DF5F-Oic-Arg 14 — M958F5bz-DArg-Arg-Aud-Ser-DF5F-Oic-Arg 18 — M960Aca-DArg-Arg-Aud-Ser-DF5F-Oic-Arg 21 — M96433Dp-DArg-Arg-Aud-Ser-DF5F-Oic-Arg 4-8 — M968Dmac-DArg-Arg-Aud-Ser-DF5F-Oic-Arg 15 — M972F5pa-DArg-Arg-Aud-Ser-DF5F-Oic-Arg — — M974 DDD-(PzO-F5F-Arg)₂ — — M976DDD-(DNiK-F5F-Arg)₂ — — M978 DDD-(DPzK-F5F-Arg)₂ — — M980DDD-(DPzO-F5F-Arg)₂ — — M1024 SUB-(DArg-Arg-Eac-Ser-DF5f-Nc7G-Arg)₂ 147.2 M1026 DTP-(DArg-Arg-Eac-Ser-DF5F-Nc7G-Arg)₂ 70 6.9 M1028SBEC-(DArg-Arg-Eac-Ser-DF5F-Nc7G-Arg)₂ 28 6.7 M1030EGS-(DArg-Arg-Eac-Ser-DF5F-Nc7G-Arg)₂ 51 7.0 M1034DDD-(DArg-F5F-DArg-NH₂)₂ — — M1036 DDD-(DArg-F5F-DArg)₂ 40 5.4 M1038ε-SUB-(Lys-DArg-Arg-Eac-Ser-DF5F-Nc7G-Arg)₂ — 6.4 M1042Aca-DArg-Arg-Eac-Ser-DF5F-Oic-Arg — — M1044Gun₂-BApg-DArg-Arg-Eac-Ser-DF5F-Oic-Arg — — M1046(F5c-DArg-Igl-Arg)₂-DDA — — Footnotes: ^(a)ED₅₀ for killing of SCLCstrain SHP-77 in vitro, μM. ^(b)pA₂ for bradykinin antagonist activityon isolated guinea pig ileum. The pD₂ of bradykinin is 7.4 on ileum.Higher numbers indicate higher potency. ^(c)pA₂ for bradykininantagonist potency on cloned human B2 receptors, pM. The pD₂ forbradykinin is 11. Higher numbers indicate higher potency. ^(d)Dataincluded for comparison Inact = inactive; Mixed = showing both agonistand antagonist activity; Wk = weak

TABLE 2 ACTIVITIES OF COMPOUNDS NOT RELATED TO BRADYKININ NUMBERSTRUCTURE MTT^(a) GPI^(b) HUMAN^(c) B9948 Arg-DNMF-DTrp-Phe-DTrp-Leu 2.8Wk B10222 DNMF-DTrp-Phe-DTrp-LeuΨ(CH₂NH)Leu-NH₂ 6.0 5.2 B10224-1α-DDD-(Lys-DNMF-DTrp-Phe-DTrp- 13 — LeuΨ(CH₂NH)Leu-NH₂)₂ B10224-2α-DDD-(Lys-DNMF-DTrp-Phe-DTrp- 7 — LeuΨ(CH₂NH)Leu-NH₂)₂ B10228DDD-(DNMF-DTrp-Phe-DTrp-LeuΨ(CH₂NH)Leu- 40 Wk NH₂)₂ B10242Arg-Pro-Lys-Pro-DTrp-Gln-DTrp-Phe-DTrp- 40 5.6 LeuΨ(CH₂NH)Leu-NH₂ B10244DArg-Arg-Pro-Lys-Pro-DTrp-Gln-DTrp-Phe- 12 5.4 DTrp-LeuΨ(CH₂NH)Leu-NH₂B10246 DArg-Pro-Lys-Pro-DTrp-Gln-DTrp-Phe-DTrp- 12 6.1LeuΨ(CH₂NH)Leu-NH₂ >278 F5c-Iqa-Atmp 9 5.3 — M8 Gun-Eac-D2Nal-PgF —Inact M12 Dcg-Igl-Aqu 20 5.0 — M18 Dcg-2Nal-Aqu 30 6.6 — M26Gun-2Nal-GaP — 4.9 M30 Dcg-2Nal-Apa — 5.4 M32 Gun-2Nal-Apa — 4.8 M36Dcg-D2Nal-Apa — 5.0 M38 Gun-D2Nal-Apa — 4.9 M62 Dcg-2Nal-Ama — 4.8 M64Dcg-2Nal-APa-Sud — Ag M72-1 Dcg-Igl-Apa — 4.7 M72-2 Dcg-Igl-APa(anisyl)— 4.6 M76 Dcg-2Nal-mABz — 4.9 M92-1 Dcg-2Nal-mA₂Bz — 5.0 M92-2Dcg-2Nal-mA₂Bz(Gun) — 4.8 M92-4 Dcg-2Nal-3Pal — 5.0 M104 Dcg-2Nal-3Pal —4.9 M112 Dcg-D2Nal-mABz — 5.1 M120 Dcg-2Nal-pABz — 4.7 M122-1Mcg-APa-mABz — 5.0 M122-2 Dcg-Apa-mABz — 4.6 M136 Sin-F5F-3Pal — InactM162 Dcg-2Nasl-Asp — 4.9 M168-1 2Nap-PaF(Mcg) — 4.8 M168-2 2Nap-PaF(Dcg)— 4.7 M172 Inp-Dpr(Dcg-2Nal) — 4.9 M174 Dcg-Asp-Aqu — Inact M180F5c-pABz-2Nal — 5.1 M188B Dcg-2Nal-Asp(Aqu) — 5.5 M202 F5c-Gly-mABz-Nal— 5.1 M204 Ac-Pac-Gly-mABz-Nal — 5.0 M218 2Nal-Atmp Inact 4.8 M222Dcg-2Nal-Atmp 15 6.8 Inact M228-2 Dcg(Me)-2Nal-Atmp(Me) 15 7.6 — M236Dcg-Igl-Atmp >50 4.7 — M240 Dcg-F5f-Atmp 32 5.1 — M244A Dcg-2Nal-Atpm 125.0 M244B Dcg-2Nal-Atpc Inact 4.9 M246 Dcg-D2Nal-Atmp >50 5.7 M248F5c-2Nal-Atmp 3.2 6.2 — M250 Aca-2Nal-Atmp — 5.2 M252 Dhq-2Nal-AtmpInact 4.8 M254 TDIM-(2Nal-Atmp)₂ 5 5.1 — M254-1 TDIM-(2Nal-Atmp)₂ 5 5.1M254-2 TDIM-(2Nal-Atmp)₂ 5 5.8 M258 Dcg-Igl-Aptp — 5.0 M262Dcg-D2Nal-Atmp 6 5.3 M264 Dcg-Trp-Atmp 16 4.7 — M266 Dcg-Apa-Atmp 42 4.8— M268 F5c-2Nal-Tpac 10 5.2 — M270 Dcg-2Nal-Tpac — 6.1 M272Dpa-2Nal-Atmp 9 5.2 — M274 Sin-2Nal-Atmp 36 4.7 — M276 Dca-2Nal-Atmp 4.65.3 — M280 TDIM-(Igl-Atmp)2 6 5.2 — M280-1 Ctim-Igl-Atmp 21 Wk M280-2TDIM-(Igl-Atmp)₂ 6 5.2 M286 Dtp-(2Nal-Atmp)₂ 24 5.1 — M288 Boc-2Nal-AtmpInact 5.2 — M288A Boc-2Nal-Atmp >85 5.2 M290-1 Btac-(2Nal-Atmp)₂ >60 WkM290-2 Btac-(2Nal-Atmp)₃ 20 Wk M292 Pac-Igl-Atmp 40 Wk M294DDD-(Pac-Igl-Atmp)₂ 1.8 Inact 10.3 M296 Pya-Bip-Atmp 15 Wk M302Atcp-2Nal-Atmp 3.5 5.2 M304 TDIM-(2Nal-Dmm)₂ 4.2 5.9 — M306Gbz-2Nal-Atmp >100 — M308 Pac-2Nal-Atmp >75 5.0 M310DDD-(Pac-2Nal-Atmp)₂ 1.2 5.1 Inact M312 Tfmc-2Nal-Atmp 3.2 5.5 — M314F5c-2Nal-Aqd 25 4.6 M316 F5c-Tyr-Atmp 50 — M318 F5c-Tyr(Bzl)-Atmp 3.65.1 — M320 F5c-Oic-Atmp 13 — — M322 F5c-Tic-Atmp 7.6 — — M324Dmac-2Nal-Atmp 3 5.2 — M336-1 Dcg-2Nal-Asp-(R,S)Aqu — 5.1 M336-2Dcg-2Nal-Asp-(R,S)Aqu — 5.4 M340 Dcg-Pac-Gly-mABz-2Nal — 4.8 M342Dcg-2Nal-Asp-Atmp — 5.4 M350 Dcg-2Nal-Glu-Atmp — 5.0 M354 Dcg-2Nal-PgF —5.2 M362 Dcg-pAPa-Asp-Atmp — 4.9 M364 F5c-pAPa-Asp-Atmp — 4.7 M368Tfmc-pAPa-Asp-Atmp — 4.7 M396 F5c-2Nal-Cys(SO₃H)-Atmp — 5.0 M408Pya-2Nal-Cyh 22 4.6 11.5 M418 F5c-BtA-Atmp 3.8 7.0 10.3 M422Pya-pABz-2Nal 52 — Inact M428 Pya-Gly-mABz-Aqd >300 — M430DDD-(BtA-Atmp)₂ 18 5.3 11.3 M432 DDD-(2Nal-Asp-Atmp)₂ 70 — 10 M436-1TDIM-BtA-Atmp 8 4.8 M436-2 TDIM-(BtA-Atmp)₂ 4.5 5.8 Ag M438F5c-3Pal-Atmp 26 4.8 10 M440 Dcg-BtA-Atmp 30 — M448 Dmac-BtA-Atmp 2.75.7 Ag M456 F5c-Cys(Meb)-Atmp 4.7 5.3 Inact M460 DDD-(3Pal-Nal-Cyh)₂ 15Wk M466 F5c-2Nal-3Ampy Inact Wk M470 F5c-2Nal-Ampz 11 5.2 Inact M472Dmac-2Nal-Ampz 25 5.3 Inact M474 Pya-2Nal-3Abza 35 Wk Inact M476Tha-BtA-Atmp 15 4.6 Inact M478 Dmac-2Nal-Thm 30 5.1 Inact M480-1HOOC-DDD-Pac-2Nal-Ampz 45 5.1 M480-2 DDD-(Pac-2Nal-Ampz)₂ — 5.6 M492F5c-mABz-2Nal-Ampz 45 5.1 Inact M506 Mse-Pac-BtA-Atmp 11 4.9 — M526F5c-2Nal-Dmp 10 5.4 Inact M536 F5c-2Nal-Dmab 4 Wk Inact M538DDD-(Pac-2-Nal-Dmp)₂ >80 Wk M568 F5po-2Nal-Atmp 10 5.8 Ag M570F5c-OC2Y-Atmp 1.8 5.6 Ag M572 Dca-2Nal-Acep 2.6 Wk Wk M574Dns-Tyr(Bzl)Atmp 4.5 — Inact M582 Dmac-OC2Y-Atmp 3 5.4 9.5 M584-ADDD-[DArg(Tos)-2Nal-Atmp]₂ 5 Inact 10.3 M584-B DDD-(DArg-2Nal-Atmp)₂ 55.7 11.3 M586-A Mse-Pac-Igl-Atmp 15 5.3 12 M586-B Seb-Pac-Igl-Atmp 40 Wk12.3 M588 α-DDD-(Lys-DArg-2Nal-Atmp)₂ 9.4 Wk 10 M592 F5c-OC2Y-Matp 1.54.9 Ag M594 F5c-MC2Y-Atmp 3.7 5.0 8 M594 F5c-MC2Y-Atmp 3.7 5.0 8 M596-ADDD-[Arg(Tos)-2Nal-Atmp]₂ 8.2 Wk Inact M596-B DDD-(Arg-2Nal-Atmp)₂ 8.2Wk Inact M602 Chc-OC2Y-Atmp 12 — 10.8 M604 Pac-2Nal-Ecap 43 4.5 M606DDD-(Pac-2Nal-Api)₂ 30 5.0 10 M614 F5c-(N-Dmb)-Tyr(Bzl)-OMe 9.1 Wk M616DDD-(Pac-1Nal-Atmp)₂ 1.4 5.4 — M618 F5c-DArg-2Nal-Arg-Matp 18 — — M620DDD-(DArg-2Nal-Arg-Matp)₂ 2.0 5.5 — M622 F5c-OC2Y-Mapp 1.2 5.7 — M624Dns-OC2Y-Matp 1.4 5.1 — M626 Pya-OC2Y-Matp 3.7 4.8 — M628 Cin-OC2Y-Matp1.6 5.2 — M630 Dmac-OC2Y-Matp 1.6 5.0 — M632 Atcp-OC2Y-Matp 1.4 5.4 —M636 DDD-(DArg-Arg-Aud-Pac-2Nal-Atmp)₂ 1.7 5.8 — M638DDD-(DArg-Igl-Arg-Matp)₂ 0.6 Inact M640 DDD-(DArg-BtA-Arg-Matp)₂ 3.0 5.9M648 F5c-PaF(Mes)-Atmp Inact 5.0 M650 Atcp-OC2Y-Mapp 3.7 — — M652Ppa-OC2Y-Mapp 7.5 5.7 — M654 Sul-Atmp Inact 4.5 M656 Sul-2Nal-Atmp 135.4 M660 DDD-(His-1Nal-Atmp)₂ 30 Wk — M662 F5c-tLeu-Atmp Inact 5.2 —M664 F5c-OCIY-Matp 1.2 5.0 M666 Dns-OCIY-Matp 1.3 5.0 M668SBEC-(DArg-2Nal-Arg-Matp)₂ 3.4 5.2 M670 DTP-(DArg-Igl-Arg-Matp)₂ Inact5.1 M672 HDD-(DArg-Igl-Arg-Matp)₂ — — M674 DDD-(DArg-F5F-Arg-Matp)₂ 3.5Wk M678 (Dns-DArg-Igl-Arg)₂-DDA 1.1 5.3 M724 F5c-DArg-Aud-OC2Y-Gly-Atmp12 5.4 M744 DDD-(DArg-2Nal-Arg-Dmab)₂ 3.4 5.3 M798 F5c-OC2Y-Dmab 37 —M800 DDD-(DArg-OC2Y-Dmab)₂ 27 5.3 M832 F5c-PFF-Dmab 47 4.6 M834DDD-(DArg-PFF-Arg-Dpea)₂ 1.6 5.3 M848 DDD-(DArg-F5F-Arg-Dmab)₂ — — M880DDD-(DArg-F5F-Arg-Dpea)₂ — — M886-1

3.2 Wk M886-2 DDD-(DArg-PFF-Arg-Dpma)₂ — Inact M892DDD-(DArg-PFF-Arg-PFF-NH₂)₂ 8.5 Wk M900 DDD-(DArg-F5F-Arg-PaF-NH₂)₂ 6.3— M916 F5c-DArg-PFF-Arg-PFF-NH₂ 5.7 4.9 M1032 DDD-(DArg-Igl-Mapp)₂ 155.4 M1040 EDTA-(OC2Y-Atmp)₄ 0.73 — Footnotes: ^(a)ED₅₀ for killing ofSCLC strain SHP-77 in vitro, μM. ^(b)pA₂ for bradykinin antagonistactivity on isolated guinea pig ileum. The pD₂ of bradykinin is 7.4 onileum. Higher numbers indicate higher potency. ^(c)pA₂ for bradykininantagonist potency on cloned human B2 receptors, pM. The pD₂ forbradykinin is 11. Higher numbers indicate higher potency. Ag = agonist;Inact = inactive; Wk = weak

TABLE 3 ACTIVITIES OF CYCLIC PEPTIDES NUMBER STRUCTURE MTT^(a) GPI^(b)B9458-2

— 6.1 B9462

7.3 6.0 B10302 c[DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg] Inact 5.2B10304 Aca-c[DArg-Arg-Pro-Hyp-Gly-Thi-Ser-Nig-Oic-Arg] Inact 6.4 B10306c[Arg-DNMF-DTrp-Phe-DTrp-Leu] Inact Wk B10312α-DDD-(c[Lys-DArg-Arg-Pro-Hyp-Gly-Thi-Ser-DF5F-Oic-Arg])₂ 3.8 — M680c[DArg-Arg-Eac-Ser-DF5F-Nc7G-Arg] — — M824 c[Ava-Igl-Ser-DF5F-Oic-Arg]Inact 5.2 M850 c[DArg-Arg-Aud-DIg1-PFF-Arg] 1.4 5.1 M868-2c[DArg-Arg-Aud-DF5F-Oic-Arg] 9.2 6.1 M870 c[DArg-Arg-Add-DF5F-Oic-Arg]5.5 5.3 M872 c[DArg-Arg-Eac-Ser-DF5F-Oic-Arg] 2.2 Inact M874c[DArg-Arg-Add-Ser-DF5F-Oic-Arg] 11   5.0 M876c[DArg-Arg-Aud-Ser-DF5F-Oic-Arg] 22.5  5.4 M878 c[DArg-ArgAdd-DIgl-PFF-Arg] 7   Wk M882 c[DArg-Arg-Add-Ser-DIgl-PFF-Arg] 4.5 InactM896 c[DArg-Arg-Eac-DIgl-PFF-Arg] 65   Wk M902c[DArg-Arg-Ava-Ser-DIgl-PFF-Arg] 30   5.5 M906c[DArg-Arg-Eac-DF5F-Oic-Arg] 45   Wk M908c[DArg-Arg-Ava-Ser-DF5F-Oic-Arg] 40   4.9 M910c[Bala-DArg-Arg-Eac-Ser-DF5F-Oic-Arg] 42   5.2 M924c[Suc-DArg-Arg-Eac-Ser-DIgl-PaF]-Arg 37   Wk M934c[DNiK-Arg-Eac-Ser-DF5F-Oic-Arg] — — M940c[DNiK-PzO-Eac-Ser-DF5F-Oic-Arg] — — M986 c[Add-DArg-F₅F-Arg] — —Footnotes: ^(a)ED₅₀ for killing of SCLC strain SHP-77 in vitro, μM.^(b)pA₂ for bradykinin antagonist activity on isolated guinea pig ileum.The pD₂ of bradykinin is 7.4 on ileum. Higher numbers indicate higherpotency. Inact = inactive; Wk = weak

TABLE 4 PREVIOUSLY DESCRIBED KNOWN PEPTIDES THAT NEWLY SHOW IN VIVOANTI-CANCER ACTIVITY NUMBER STRUCTURE B9430DArg—Arg—Pro—Hyp—Gly—Igl—Ser—DIgl—Oic—Arg B9330DArg—Arg—Pro—Hyp—Gly—Thi—Ser—DTic—Nig—Arg B10044DArg—Arg—Pro—Hyp—Gly—Igl—Ser—DF5F—Oic—Arg B10050Lys—Lys—Arg—Pro—Hyp—Gly—Igl—Ser—DTic—ChG B10206DArg—Arg—Pro—Hyp—Gly—Igl—Ser—DF5F—Nc7G—Arg B10288DArg—Arg—Pro—Hyp—Gly—Thi—Ser—DTic—Oic—Arg

These compounds showed anti-tumor activity in vivo when tested by theprocedure of Example XXI.

TABLE 5 CYTOTOXICITY IN VITRO AGAINST STRAINS OF PROSTATE CANCER COM-POUND PROSTATE CANCER CELL LINE SCLC NUMBER DU14 TSU LNCa PC-3 PPC1SHP-77 B9870 0.08 6.5 3.7 3.2 4.3 0.15 M570 1.2 2.8 3.0 1.6 3.0 1.8 M5900.01 7.0 7.0 6.3 12 4.5

Numbers are ED₅₀ (μM) for cytotoxic activity. Activity against SCLCstrain SHP-77 is included for comparison.

TABLE 6 ABBREVIATIONS USED FOR COMPOUNDS B9430 =DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic-Arg B9870 =SUIM-(DArg-Arg-Pro-Hyp-Gly-Igl-SAer-DIgl-Oic-Arg)₂ Aaa =1-Adamantaneacetyl AAA = amino acid analysis ABz = Aminobenzoic acidABza = Aminobenzyl alcohol Ac = Acetyl Ac3c =1-Amino-1-cyclopropanecarboxylic acid Ac5c =1-Amino-1-cyclopentanecarboxylic acid (“cyclo-Leu”) Ac6c =1-Aminocyclohexanecarboxylic acid Aca = 1-Adamantanecarboxyl Acep =4-Amino-1-carbethoxymethyl-2,2,6,6-tetramethylpiperidine ADA =1,3-Adamantanediacetyl Add = 12-Aminododecanoic acid Aib =α-Aminoisobutyric acid Aic = 2-Aminoindane-2-carboxylic acid A1G =α-Allylglycine (2-amino-5-pentenoic acid) Ama = Aminomethylanthranilicacid Amb = Aminomethylbenzoic acid Ampy = 3-Aminomethylpyridine Ampz =1-Amino-4-methylpiperazine Ana = Anthranilic acid APa =p-Aminophenylacetic acid APb = p-Aminophenylbutyric acid ApC =S-3-Aminopropylcysteine Api = 4-Aminopiperidine Apmp =4-Amino-1,2,2,6,6-pentamethylpiperidine Aptp =4-Amino-1-phenylmethyl-2,2,6,6-tetramethylpiperidine Aqd =4-Aminoquinaldine Aqu = 3-Aminoquinuclidine Arg(NO₂) = Arginine(Nitro)Atc = 2-Aminotetralin-2-carboxylic acid Atcp =4-Amino-3,5,6-trichlorpicolinic acid Atmp =4-Amino-2,2,6,6-tetramethylpiperidine AtmpO =4-Amino-2,2,6,6-tetramethylpiperidinyloxy Atpc =4-Amino-2,2,6,6-tetramethyl-4-piperidinecarboxylic acid Atpm =4-Amino-4-methoxycarbonyl-2,2,6,6-Tetramethylpiperidine(4-Amino-2,2,6,6-tetramethyl-4-piperidinecarboxylic acid methyl ester)Aud = 11-Aminoundecanoic acid Ava = 5-Aminovaleric acid Azt =Azetidine-2-carboxylic acid BAla = β-Alanine BApG =N,N-bis(3-aminopropyl)-glycine BAPTA =1,2-bis(2-Aminophenoxy)ethane-N,N,N′,N′-tetraacetyl Bip =Biphenylalanine Boc = (tert-Butoxycarbonyl);[(1,1-dimethylethoxy)carbonyl] BOP =Benzotriazoyloxytris(dimethylamino)phosphonium hexafluorophosphate BPHD= N,N′-bis(2,2,6,6-tetramethyl-4-piperidinyl)-1,6-hexanediamine BSH =1,6-Bissuccinimidohexane BtA = 3-Benzothienylalanine BTAC =Benzene-1,3,5-tris-carboxyamido-6-caproyl BTC = 1,3,5-BenzenetricarboxylBz = Benzoyl Bzl = Benzyl CAcH = cis-2-Amino-1-cyclohexanecarboxylicacid Cca; 2-Chlorocinnamic acid CDF = p-Chloro-D-phenylalanine ChA =α-Cyclohexylalanine Chc = α-Cyano-4-hydroxycinnamoyl ChG =α-Cyclohexylglycine CHO = Chinese hamster ovary CHTC =1,3,5-Cyclohexanetricarboxyl CHyp = cis-4-Hydroxy-proline Cin =Cinnamoyl CMeb = S-(4-Methylbenzyl cysteine CmF = (Z)p-Chloro-2,3-methanophenylalanine Cmp = 4-Carboxymethylpiperazine CpA =α-Cyclopropylalanine CpG = α-Cyclopentylglycine CpGΨ(CH₂N)Arg = CpGpseudo(CH₂NH) Arg CPTA =trans-1,2-Diaminocyclohexane-N,N,N′,N′-tetraacetyl CTAC =Cyclohexane-1,3,5-tris-carbamido-ε-caproyl Ctim =13-Carboxytridecanimidyl Cyh = Cyclohexylamine Dabz = Diaminobenzoicacid DArg(NO₂) = Nitro-Arginine Dca = Dicyclohexylacetyl Dcg =N,N′-Dicylcohexylguanidyl DCM = Dichloromethane DDA = 1,10-DecanediamineDDD = Dodecanedioyl- DDS = 2-Dodecen-1-ylsuccinyl DEA =N,N′-Diethylethylenediamine DhP = 3,4-Dehydroproline Dhq =2,3-Dehydroquinuclidine-3-carboxyl DIC =Decahydroisoquinuclidine-3-carboxyl DIEA = Diisopropylethylamine Dmab =4-Dimethylaminobenzylamine Dmac = 4-Dimethylaminocinnamyoyl Dmb =4-(Dimethylamino)benzyl DmF = 2,4-Dimethylphenylalanine DMF = Dimethylformamide DmK = ε-Dimethyllysine Dmm = 2,6-Dimethylmorpholine Dmp =3-Dimethylaminopropylamine DmtP = 5,5-Dimethyl-4-thiaproline Dns =Dansyl (5-dimethylamino-1-naphthalenesulfonyl) 22Dp =2,2-Diphenylpropionyl 33Dp = 3,3-Diphenylpropionyl Dpa = DiphenylacetylDpea = Diphenylethylamine Dpma = Diphenylmethylamine Dpr =2,3-Diaminopropionic acid DTP = Dithiobis-propionyl DTPA =Diethylenetriaminepentaacetyl Eac = ε-Aminocaproic acid Ecap =N-Ethoxycarbonyl-4-amino-piperidine (Ethyl 4-amino-1-piperidinecarboxylate) EDA = 4,4′-Ethylenedianiline EDP =4,4′-Ethylenedipiperidine EDTA = Ethylenediaminetetraacetyl EDTP =Ethylenediaminetetrapropionic acid EGS = Ethylene glcyol-bis-succinylEGTA = Ethylene glycol-bis(βaminoethyl ether)-N,N,N′,N′-tetraacetyl EOPC= 1,1′-Ethylenebis(5-oxo-3-pyrrolidinecarboxyl) ETTA =2,2′,2″,2′″-[Ethanediylidenetetrakis(thio)tetrakisacetyl F2F =Difluorophenylalanine F5bz = Pentafluorobenzoyl F5c =2,3,4,5,6-Pentafluorcinnamoyl F5F = Pentafluorophenylalanine F5pa =2,3,4,5,6-Pentafluorocinnamoyl F5po = 2,3,4,5,6-PentafluorophenoxyacetylFbz = para-Fluorobenzoyl Flu = Fluorescein thiourea Gaa =Guanidinoacetyl GaP = 2-Guanidyl-3-(4-aminophenyl)propionic acid Gbz =4-Guanidinobenzoyl Glt = Glutaryl Gun = Guanidyl HATU =O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate HbQ = 4-Hydroxybutylglutamine HDA =1,6-Hexanediamine HDD = Hexadecanedioyl HF = Hydrogen fluoride HFG =hexafluoroglutaroyl HiG = Hexahydro-2-indanylglycine HOAt =1-Hydroxy-7-azabenzotriazole hPhe = Homo-phenylalanine HPLC = highperformance liquid chromatography Hxa = Hexanoic acid Hyp =trans-4-Hydroxyproline Ica = Indoline-2-carboxylic acid Igl =α-2-Indanylglycine Ing = α-1-Indanylglycine Inp = Isonipecotic acid Iq2a= 6,7-Dimethoxy-3,4-Dihydro-1-isoquinolineacetic Acid Iq4a =6,7-Dimethoxy-1,2,3,4-tetrahydro-1-isoquinolineacetic acid Lau = LauroylLeu(r)Leu = Leu-pseudo(CH₂NH)Leu LeuΨ(CH₂NH)Leu = Leu-pseudo(CH₂NH)LeuLDMS = laser desorption mass spectrometry mA₂Bz = 3,5-Diaminobenzoicacid MaG = α-Methallylglycine (2-amino-3-methyl-4-pentenoic acid) Mapp =4-(Methylamino)-1,2,2,6,6-pentamethylpiperidine Matp =4-(Methylamino)-2,2,6,6-tetramethylpiperidine MatpO =4-(N-methylamino)-2,2,6,6-tetramethylpiperidinyloxy MBC =S-(4-methylbenzylcysteine MBHA = Methylbenzhydrylamine MC2Y =N-Methyl-O-2,6-dichlorobenzyl-tyrosine Mca = 2-Methylcinnamic acid Mcg =Monocyclohexylguanidyl Meb = Methylbenzyl MeP = 2,4-Methanoproline Mes =Methanesulfonyl MFE = (E)-2,3-Methanophenylalanine MFF =meta-Fluorophenylalanine Mosi = Methoxy-suberimido Moti =14-Methoxytetradecanediimidoyl Mse = Methoxysebacyl MTT =(3-(4,5)-Dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide Nal =β-Naphthylalanine Nap = Naphthoyl Nba = Norbornane-2-acetyl Nbc =Norbornenedicarboxyl Nbi = Norbornenedicarboximide Nbn =2-Aminonorbornane-2-carboxylic acid Nc5G = N-Cyclopentylglycine Nc6G =N-Cyclohexylglycine Nc7G = N-Cycloheptylglycine Nc8G =N-Cyclooctylglycine Nig = N-2-Indanylglycine NiK = ε-NicotinoyllysineNMF = N-Methylphenylalanine NSCLC = non-small cell carcinoma OBS =O-Benzylserine OBT = O-Benzylthreonine OBY = O-Benzyltyrosine OC2Y =O-2,6-Dichlorobenzyltyrosine OCIY =O-2,6-Dichlorobenzyl-3,5-diiodotyrosoine Oct = Octanoyl Oic =Octahydroindole-2-carboxylic acid OMe = O-Methyl OMY = O-MethyltyrosineOSY = Tyrosine O-sulfate ester Paa = Phenylacetyl Pac = 4-Aminocinnamicacid PaF = p-Aminophenylalanine Pal = β-Pyridylalanine Pba =Phenylbutyryl Pcc = trans-2-Phenyl-1-cyclopropanecarboxylic acid PCF =p-Chlorphenylalanine Pcpa = α-Phenylcyclopentaneacetyl PdF =p-Dicyclohexylguanidylphenylalanine PFF = p-Fluorophenylalanine PFS =Perfluorosuberoyl PgF = p-Guanidinophenylalanine PheOL = PhenylalaninolPhG = Phenylglycine Pip = Pipecolic acid (“homo-Pro”) PipA =β-3-Piperidylalanine PNF = p-Nitrophenylalanine Ppa = PhenylpropionylPya = trans-3-(3-Pyridyl)acryloyl PyAOP =7-Azabenzotriazol-1-yloxytris(pyrrolidino)phosphoniumhexafluorophosphate PzK = ε-Pyrazinoyllysine PzO = 4-PyrazinoylornithineSaa = trans-Styrylacetic acid SBEC = Sulfo-bis-ethoxycarbonyl Sbl =Sebacoyl SCLC = small cell lung carcinoma Seb = Sebacyl Sin = Sinapinyl(3,5-dimethoxy-4-hydroxycinnamoyl-) Ste = Stearoyl Sua = Sulfanilamide(4-Aminobenzenesulfonamide) SUB = Suberyl Suc = Succinyl Sud =Sulfadiazine SUIM = Suberimidyl Sul = Sulindac Tba = t-Butyl-acetyl TDIM= Tetradecanediimidyl TEA = Triethylamine TFA = Trifluoroacetic acidTfmc = trans-4-(Trifluoromethyl)cinnamoyl Tha = 3-(2-Thienyl)acryloylThi = β-2-Thienylalanine Thm = Thiomorpholine Thz =Thiazolidine-4-carboxylic acid (4-thiaproline) Tic =1,2,3,4-Tetrahydroisoquinoline-3-carboxylic acid TLC = thin layerchromatography TLeu = tert-Leucine TMF = 2,4,6-TrimethylphenylalanineTos = p-Toluenesulfonyl Tpac =2,2,5,5-Tetramethyl-3-(aminoethyl)-pyrroline-3-carboxamide TREN =tris(2-Aminoethyl)amine Trx = Tranexamic acid(trans-4-((Aminomethyl))cyclohexanecarboxylic acid)

1 1 9 PRT Homo sapiens 1 Arg Pro Pro Gly Phe Ser Pro Phe Arg 1 5

What is claimed is:
 1. The compound of structure:2,3,4,5,6-Pentafluorocinnamoyl-D-Arginine-Arginine-Proline-Trans-4-hydroxyproline-Glycine-α-2-Indanylglycine-Serine-D-α-2-Indanylglycine-Octahydroindole-2-carboxylicacid-Arginine or a pharmaceutically acceptable salt thereof.
 2. Thecompound of the structure:2,3,4,5,6Pentafluorocinnamoyl-Lysine-Lysine-Arginine-Proline-Trans-4-hydoxyproline-Glycine-α-Cylcopentylglycine-Serine-D-1,2,3,4-Tetrahydroisoquinoline-3-carboxylicacid-α-Cylcopentylglycine or a pharmaceutically acceptable salt thereof.3. The compound of structure:2,3,4,5,6-Pentafluorocinnamoyl-D-ε-Nicotinoyllysine-4-(2-pyrazinecarboxyl)ornithineProline-Trans-4-hydroxyproline-Glycine-α-2-Indanylglycine-Serine-D-α-2-Indanylglycine-Octahydroindole-2-carboxylicacid-Arginine or a pharmaceutically acceptable salt thereof.
 4. Thecompound of structure:2,3,4,5,6-Pentafluorocinnamoyl-p-Fluorophenylalanine-Arginine or apharmaceutically acceptable salt thereof.
 5. The compound comprising thestructure: Dodecanedioyl-(D-Arginine-Pentaflurorphenylalanine-Arginine)₂or a pharmaceutically acceptable salt thereof.
 6. The compound ofstructure:2,3,4,5,6-Pentafluorocinnamoyl-O-2,6-Dichlorobenzyltyrosine-4-Amino-2,2,6,6-tetramethylpiperidineor a pharmaceutically acceptable salt thereof.
 7. The compound ofstructure:Dodecanedioyl-(D-Arginine-α-2-Indanylglycine-Arginine-4-(Methylamino)-2,2,6,6-tetramethylpiperidine)₂or a pharmaceutically acceptable salt thereof.
 8. The compoundcomprising the structure:Ethylenediaminetetraacetyl-(O-2,6-Dichlorobenzyltyrosine-4-(Methylamino)-2,2,6,6-tetramethylpiperidine)₄or a pharmaceutically acceptable salt thereof.
 9. The compound ofstructure:(5-dimethylamino-1-naphthalenesulfonyl-D-Arginine-α-2-Indanylglycine-Arginine)₂-1,10-Decanediameor a pharmaceutically acceptable salt thereof.
 10. The compound ofstructure:Dodecanedioyl-(D-Arginine-β-2Naphthylalanine-Arginine-4-(methylamino)-2,2,6,6-tetramethylpiperidine)₂or a pharmaceutically acceptable salt thereof.
 11. The compound ofstructure: Decanedioyl-(-4-Aminocinnamicacid-α-2-Indanylglycine-4-amino-2,2,6,6-tetramethylpiperidine),(-Lysine-D-Arginine-Arginine-Proline-Trans-4-hydroxyproline-Glycine-α-2-Indanylglycine-Serine-D-α-2-Indanylglycine-Octrahydroindole-2-carboxylicacid-Arginine), or a pharmaceutically acceptable salt thereof.